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Literature summary for 1.14.20.1 extracted from
- Iterative combinatorial mutagenesis as an effective strategy for generation of deacetoxycephalosporin C synthase with improved activity toward penicillin G (2012), Appl. Environ. Microbiol., 78, 7809-7812.
Protein Variants
| Protein Variants | Comment | Organism |
|---|---|---|
| C155Y | site-directed mutagenesis | Streptomyces clavuligerus |
| C155Y/Y184H/V275I/C281Y | site-directed mutagenesis, the mutant shows increased activity with penicillin G compared to the wild-type enzyme | Streptomyces clavuligerus |
| C281Y | site-directed mutagenesis | Streptomyces clavuligerus |
| I305L | site-directed mutagenesis, the mutant shows increased activity with penicillin G compared to the wild-type enzyme | Streptomyces clavuligerus |
| I305M | site-directed mutagenesis, the mutant shows increased activity with penicillin G compared to the wild-type enzyme | Streptomyces clavuligerus |
| I305M/S261M | site-directed mutagenesis, the mutant shows increased activity with penicillin G compared to the wild-type enzyme | Streptomyces clavuligerus |
| I305M/T213V | site-directed mutagenesis, the mutant shows increased activity with penicillin G compared to the wild-type enzyme | Streptomyces clavuligerus |
| I305M/T213V/M73T | site-directed mutagenesis, the mutant shows increased activity with penicillin G compared to the wild-type enzyme | Streptomyces clavuligerus |
| I305M/T213V/S261M | site-directed mutagenesis, the mutant shows increased activity with penicillin G compared to the wild-type enzyme | Streptomyces clavuligerus |
| M73T | site-directed mutagenesis | Streptomyces clavuligerus |
| additional information | since the natural substrate of the enzyme, penicillin N, is not available in large quantities, engineering of a scDAOCS capable of converting the cheap and easily available substrate penicillin G is valuable for large-scale production of 7-aminodeacetoxycephalosporanic acid, 7-ADCA | Streptomyces clavuligerus |
| N304K | site-directed mutagenesis | Streptomyces clavuligerus |
| Q126M | site-directed mutagenesis | Streptomyces clavuligerus |
| S261A | site-directed mutagenesis | Streptomyces clavuligerus |
| S261M | site-directed mutagenesis | Streptomyces clavuligerus |
| T213V | site-directed mutagenesis | Streptomyces clavuligerus |
| V275I | site-directed mutagenesis | Streptomyces clavuligerus |
| Y184H | site-directed mutagenesis | Streptomyces clavuligerus |
KM Value [mM]
| KM Value [mM] | KM Value Maximum [mM] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 1.6 | - |
penicillin G | mutant C155Y/Y184H/V275I/C281Y, pH and temperature not specified in the publication | Streptomyces clavuligerus |  |
| 1.62 | - |
penicillin G | mutant I305M/S261M, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 1.92 | - |
penicillin G | mutantI305M/T213V/M73T, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 2.1 | - |
penicillin G | mutant I305L, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 2.49 | - |
penicillin G | mutant I305M/T213V/S261M, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 3.22 | - |
penicillin G | mutant I305M/T213V, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 3.58 | - |
penicillin G | mutant I305M, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 14.38 | - |
penicillin G | wild-type enzyme, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
Natural Substrates/ Products (Substrates)
| Natural Substrates | Organism | Comment (Nat. Sub.) | Natural Products | Comment (Nat. Pro.) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| penicillin N + 2-oxoglutarate + O2 | Streptomyces clavuligerus | the enzyme catalyzes the ring expansion of penicillin substrates into a ring-expanded product to produce cephamycin C | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
Organism
| Organism | UniProt | Comment | Textmining |
|---|---|---|---|
| Streptomyces clavuligerus | - |
- |
- |
Substrates and Products (Substrate)
| Substrates | Comment Substrates | Organism | Products | Comment (Products) | Rev. | Reac. |
|---|---|---|---|---|---|---|
| penicillin G + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | phenylacetyl-7-aminodeacetoxycephalosporanic acid + succinate + CO2 + H2O | - |
? | |
| penicillin N + 2-oxoglutarate + O2 | - |
Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
| penicillin N + 2-oxoglutarate + O2 | the enzyme catalyzes the ring expansion of penicillin substrates into a ring-expanded product to produce cephamycin C | Streptomyces clavuligerus | deacetoxycephalosporin C + succinate + CO2 + H2O | - |
? | |
Synonyms
| Synonyms | Comment | Organism |
|---|---|---|
| DAOCS | - |
Streptomyces clavuligerus |
Turnover Number [1/s]
| Turnover Number Minimum [1/s] | Turnover Number Maximum [1/s] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.054 | - |
penicillin G | wild-type enzyme, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.23 | - |
penicillin G | mutant C155Y/Y184H/V275I/C281Y, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.37 | - |
penicillin G | mutant I305L, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.537 | - |
penicillin G | mutant I305M/S261M, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.589 | - |
penicillin G | mutantI305M/T213V/M73T, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.65 | - |
penicillin G | mutant I305M, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.654 | - |
penicillin G | mutant I305M/T213V, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.668 | - |
penicillin G | mutant I305M/T213V/S261M, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
kcat/KM [mM/s]
| kcat/KM Value [1/mMs-1] | kcat/KM Value Maximum [1/mMs-1] | Substrate | Comment | Organism | Structure |
|---|---|---|---|---|---|
| 0.004 | - |
penicillin G | wild-type enzyme, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.144 | - |
penicillin G | mutant C155Y/Y184H/V275I/C281Y, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.176 | - |
penicillin G | mutant I305L, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.182 | - |
penicillin G | mutant I305M, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.203 | - |
penicillin G | mutant I305M/T213V, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.268 | - |
penicillin G | mutant I305M/T213V/S261M, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.307 | - |
penicillin G | mutantI305M/T213V/M73T, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
| 0.332 | - |
penicillin G | mutant I305M/S261M, pH and temperature not specified in the publication | Streptomyces clavuligerus | |
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